1887

Abstract

The influenza virus matrix protein (M1) possesses a cysteine and histidine (CCHH) motif in the helix 9 (H9) and adjacent region ( ATEQIADSQRS ). The CCHH motif has been proposed as a putative zinc finger motif and zinc-binding activity has been implicated in virus uncoating as well as transcription inhibition and mRNA regulation. The function of the CCHH motif in the influenza virus life cycle was investigated by site-directed mutagenesis (alanine replacement) and by rescuing mutant viruses by reverse genetics. Mutant viruses containing an alanine replacement of the cysteine and histidine residues, either individually or in combination, were seen to exhibit wt phenotype in multiple virus growth cycles and plaque morphology. In addition, synthetic peptides containing the putative zinc finger motif did not inhibit virus replication in MDCK cells. However, mutation of Ala in H9 was lethal for rescuing infectious virus. These data show that the CCHH motif does not provide a critical function in the influenza virus life cycle in cell culture and that the zinc-binding function may not be involved in virus biology. However, the lethal phenotype of the Ala mutation shows that the H9 region of M1 provides some other critical function(s) in virus replication.

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2003-11-01
2024-03-28
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References

  1. Arzt S., Baudin F., Barge A., Timmins P., Burmeister W. P., Ruigrok R. W. H. 2001; Combined results from solution studies on intact influenza virus M1 protein and from a new crystal form of its N-terminal domain show that M1 is an elongated monomer. Virology 279:439–446
    [Google Scholar]
  2. Elster C., Fourest E., Baudin F., Larsen K., Cusack S., Ruigrok W. H. 1994; A small percentage of influenza virus M1 protein contains zinc but zinc does not influence in vitro M1–RNA interaction. J Gen Virol 75:37–42
    [Google Scholar]
  3. Elster C., Larsen K., Gagnon J., Ruigrok R. W. H., Baudin F. 1997; Influenza virus M1 protein binds to RNA through its nuclear localization signal. J Gen Virol 78:1589–1596
    [Google Scholar]
  4. Fernandez-Pol J. A., Hamilton P. D., Klos D. J. 2001; Essential viral and cellular zinc and iron containing metalloproteins as targets for novel antiviral and anticancer agents: implications for prevention and therapy of viral diseases and cancer. Anticancer Res 21:931–957
    [Google Scholar]
  5. Harris A., Sha B., Luo M. 1999; Structural similarities between influenza virus matrix protein M1 and human immunodeficiency virus matrix and capsid proteins: an evolutionary link between negative-stranded RNA viruses and retroviruses. J Gen Virol 80:863–869
    [Google Scholar]
  6. Harris A., Forouhar F., Qiu S., Sha B., Luo M. 2001; The crystal structure of the influenza matrix protein M1 at neutral pH: M1–M1 protein interfaces can rotate in the oligomeric structures of M1. Virology 289:34–44
    [Google Scholar]
  7. Hoffmann E., Neumann G., Kawaoka Y., Hobom G., Webster R. G. 2000; A DNA transfection system for generation of influenza A virus from eight plasmids. Proc Natl Acad Sci U S A 97:6108–6113
    [Google Scholar]
  8. Hui E. K.-W., Nayak D. P. 2001; Role of ATP in influenza virus budding. Virology 290:329–341
    [Google Scholar]
  9. Hui E. K.-W., Nayak D. P. 2002; Role of G protein and protein kinase signalling in influenza virus budding in MDCK cells. J Gen Virol 83:3055–3066
    [Google Scholar]
  10. Hui E. K.-W., Yang Y. H., Yung B. Y.-M. 1992; Schedule-dependent sphinganine potentiation of retinoic acid-induced differentiation, cell growth inhibition, and nucleophosmin translocation in a human leukemia cell line (HL-60). Exp Hematol 20:454–461
    [Google Scholar]
  11. Hui E. K.-W., Barman S., Yang T. Y., Nayak D. P. 2003; Basic residues of the helix six domain of influenza virus M1 involved in nuclear translocation of M1 can be replaced by PTAP and YPDL late assembly domain motifs. J Virol 77:7078–7092
    [Google Scholar]
  12. Iuchi S. 2001; Three classes of C2H2 zinc finger proteins. Cell Mol Life Sci 58:625–635
    [Google Scholar]
  13. Judd A. K., Sanchez A., Kharitonenkov I., Moscona A., Nasser E., Bucher D. J. 1992; M-protein peptides of influenza virus: application as antiviral agents. In Peptides pp  694–696 Edited by Smith J. A., Rivier J. E. Leiden: ESCOM;
    [Google Scholar]
  14. Judd A. K., Sanchez A., Bucher D. J., Huffman J. H., Bailey K., Sidwell R. W. 1997; In vivo anti-influenza virus activity of a zinc finger peptide. Antimicrob Agents Chemother 41:687–692
    [Google Scholar]
  15. Lamb R. A., Krug R. M. 2001; Orthomyxoviridae : the viruses and their replication. In Fields Virology pp  725–769 Edited by Knipe D. M., Howley P. M. Philadelphia: Lippincott Williams & Wilkins;
    [Google Scholar]
  16. Lemay G., Danis C. 1994; Reovirus λ 1 protein: affinity for double-stranded nucleic acids by a small amino-terminal region of the protein independent from the zinc finger motif. J Gen Virol 75:3261–3266
    [Google Scholar]
  17. Liu T., Ye Z. 2002; Restriction of viral replication by mutation of the influenza virus matrix protein. J Virol 76:13055–13061
    [Google Scholar]
  18. Matthews J. M., Kowalski K., Liew C. K., Sharpe B. K., Fox A. H., Crossley M., MacKay J. P. 2000; A class of zinc finger involved in protein–protein interactions: biophysical characterization of CCHC fingers from Fog and U-shaped. Eur J Biochem 267:1030–1038
    [Google Scholar]
  19. Nasser E. H., Judd A. K., Sanchez A., Anastasiou D., Bucher D. J. 1996; Antiviral activity of influenza virus M1 zinc finger peptides. J Virol 70:8639–8644
    [Google Scholar]
  20. Nayak D. P., Hui E. K.-W. 2002; Assembly and morphogenesis of influenza viruses. Recent Res Dev Virol 4:35–54
    [Google Scholar]
  21. Neumann G., Whitt M. A., Kawaoka Y. 2002; A decade after the generation of a negative-sense RNA virus from cloned cDNA – what have we learned?. J Gen Virol 83:2635–2662
    [Google Scholar]
  22. Okada A., Miura T., Takeuchi H. 2003; Zinc- and pH-dependent conformational transition in a putative interdomain linker region of the influenza virus matrix protein M1. Biochemistry 42:1978–1984
    [Google Scholar]
  23. Perez D. R., Donis R. O. 1998; The matrix 1 protein of influenza A virus inhibits the transcriptase activity of a model influenza reporter genome in vivo . Virology 249:52–61
    [Google Scholar]
  24. Pieler T., Bellefroid E. 1994; Perspectives on zinc finger protein function and evolution: an update. Mol Biol Rep 20:1–8
    [Google Scholar]
  25. Rey O., Nayak D. P. 1992; Nuclear retention of M1 protein in a temperature-sensitive mutant of influenza (A/WSN/33) virus does not affect nuclear export of viral ribonucleoproteins. J Virol 66:5815–5824
    [Google Scholar]
  26. Sha B., Luo M. 1997; Structure of a bifunctional membrane-RNA binding protein, influenza virus matrix protein M1. Nat Struct Biol 4:239–244
    [Google Scholar]
  27. Shishkov A. V., Goldanskii V. I., Baratova L. A., Fedorova N. V., Ksenofontov A. L., Zhirnov O. P., Galkin A. V. 1999; The in situ spatial arrangement of the influenza A virus matrix protein M1 assessed by tritium bombardment. Proc Natl Acad Sci U S A 96:7827–7830
    [Google Scholar]
  28. Takatsuji H. 1998; Zinc finger transcription factors in plants. Cell Mol Life Sci 54:582–596
    [Google Scholar]
  29. Wakefield L., Brownlee G. G. 1989; RNA-binding properties of influenza A virus matrix protein M1. Nucleic Acids Res 17:8569–8580
    [Google Scholar]
  30. Watanabe K., Handa H., Mizumoto K., Nagata K. 1996; Mechanism for inhibition of influenza virus RNA polymerase activity by matrix protein. J Virol 70:241–247
    [Google Scholar]
  31. Whittaker G., Kemler I., Helenius A. 1995; Hyperphosphorylation of mutant influenza virus matrix protein, M1, causes its retention in the nucleus. J Virol 69:439–445
    [Google Scholar]
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